Survivin-短发夹RNA抑制结肠癌SW480细胞生长的研究

目的 探讨靶向survivin的短发夹RNA(shRNA)对人结肠癌SW480细胞的基因下调效应和细胞生长的抑制作用.方法 构建靶向survivin的shRNA质粒载体,然后转染入结肠癌SW480细胞.用逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western Blot)检测survivin的mRNA和蛋白的表达情况,MTT实验评价转染后各组细胞的增殖情况.结果 靶向survivin的shRNA能够明显下调survivin mRNA和蛋白质表达(P＜0.05),其抑制率分别是41.49%和40.82%.MTT实验提示转染了特异性靶向survivin shRNA的SW480细胞在转染48 h后开始呈现随着时间延长而进行性生长抑制的现象.在转染后72 h该细胞存活率仅为43.6%,与空白对照组相比有显著性的降低(P＜0.05).结论 靶向survivin的特异性shRNA对结肠癌SW480细胞能够明显地下调survivin基因表达,并且显著抑制细胞的增殖.Abstract：Objective To observe the inhibitory effect of survivin gene expression and cell proliferation of the human colon cancer cell line SW480 caused by surviving-shRNA. Methods The survivin-shRNA was constructed and transfected into SW480 cells with LipofectamineTM2000. The down-regulations of survivin expressions were detected by RT-PCR and western blot analysis. The cell proliferation inhibitions were determined by MTT assay. Results The survivin-shRNA successfully and efficiently suppressed the expression of survivin mRNA and protein(P＜0.05).The expression inhibition rates were 41.49% and 40.82% at the mRNA and protein level respectively. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates on a time-dependent manner since 48 hours post-transfection. The cell viability of SUR group was 47.8% at 72 h,which was of statistical significance compared with that of blank control group (P＜0.05).Conclusions The specific survivin-shRNA could down-regulate the expression of survivin gene and inhibit the growth of SW480 cells.

The study of survivin-shRNA inhibits growth of human colon cancer SW480 cells